大黄酸调节Ras/ERK信号通路对肝细胞癌细胞增殖、迁移和侵袭的影响

Effects of Rhein on Proliferation,Migration and Invasion of Hepatocellular Carcinoma Cells by Regulating Ras/ERK Signaling Pathway

目的:探讨大黄酸调节大鼠肉瘤蛋白(Ras)/胞外信号调控激酶(ERK)信号通路对肝细胞癌(HCC)细胞增殖、迁移和侵袭的影响。方法:使用不同浓度(0、12.50、25、50、100、150、200 mol/L)大黄酸处理HepG2细胞,检测细胞活性,筛选最佳大黄酸浓度。将细胞分为对照组、大黄酸低、中、高浓度组、大黄酸高浓度+Ras/ERK激活剂组(大黄酸高浓度+ML-099组),分别检测各组细胞集落形成数、划痕愈合率、细胞侵袭数和Ras、p-ERK、ERK蛋白表达。结果:大黄酸以浓度和时间依赖性降低HepG2细胞活性(P<0.05),选用25、50、100 mol/L处理HepG2细胞24 h用于后续实验;与对照组比较,大黄酸低、中、高浓度组细胞集落形成数、G0/G1细胞比例、细胞划痕愈合率、细胞侵袭数和原癌基因(c-Myc)、细胞周期蛋白D1(CyclinD1)、Ras、p-ERK/ERK蛋白表达呈浓度依赖性降低,S期和G2/M细胞比例、p53蛋白表达呈浓度依赖性增加(P<0.05);与大黄酸高浓度组比较,大黄酸高浓度+ML-099组细胞集落形成数、G0/G1细胞比例、细胞划痕愈合率、细胞侵袭数和c-Myc、CyclinD1、Ras、p-ERK/ERK蛋白表达显著增加,S期和G2/M细胞比例、p53蛋白表达显著降低(P<0.05)。结论:大黄酸可能通过抑制Ras/ERK信号通路抑制HCC细胞增殖、迁移和侵袭。

Objective:To investigate the effects of rhein on the proliferation,migration and invasion of hepatocellular carcinoma(HCC)cells by regulating rat sarcoma protein(Ras)/extracellular signal-regulated kinase(ERK)signaling pathway.Methods:HepG2 cells were treated with different concentrations(0,12.50,25,50,100,150,200 mol/L)of rhein to detect cell activity and screen the best concentration of rhein.The cells were divided into control group,low,medium and high concentration rhein group,high concentration rhein+Ras/ERK activator group(high concentration rhein+ML-099 group),the number of cell colony formation,scratch healing rate,the number of cell invasion and the expression of Ras,p-ERK and ERK protein were detected in each group.Results:Rhein reduced the activity of HepG2 cells in a concentration-and time-dependent manner(P<0.05),HepG2 cells were treated with 25,50,100 mol/L for 24 h for subsequent experiments.Compared with control group,the number of cell colony formation,the proportion of G0/G1 cells,cell scratch healing rate,the number of cell invasion and the expression of proto-oncogene(c-Myc),cyclin D1(CyclinD1),Ras and p-ERK/ERK protein in low,medium and high concentration groups of rhein decreased in a concentration-dependent manner,and the proportion of S phase and G2/M cells and the expression of p53 protein increased in a concentration-dependent manner(P<0.05).Compared with rhein high concentration group,the number of cell colony formation,the proportion of G0/G1 cells,cell scratch healing rate,the number of cell invasion and the expression of c-Myc,CyclinD1,Ras and p-ERK/ERK protein in rhein high concentration+ML-099 group were significantly increased,and the proportion of S phase and G2/M cells and the expression of p53 protein were significantly decreased(P<0.05).Conclusion:Rhein may inhibit the proliferation,migration and invasion of HCC cells by inhibiting the Ras/ERK signaling pathway.